• Journal of fish pathology
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• v.35 no.1
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• pp.113-120
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• 2022

Scuticociliatosis caused by Miamiensis avidus is a very important parasitic disease in olive flounder farming industry. The aim of this study was to determine effect of combined treatment with blue LED (light-emitting diode) illumination and formalin on olive flounder (Paralichthys olivaceus) infected with M. avidus. Different intensity of 405 nm LED (20, 40, and 60 μmol·m^-2·s^-1) was illuminated on 2.2×10⁴ cells/well of M. avidus in a 24 well microplate for 24 h. Also, 2.4×10⁴ cells/well of M. avidus were exposed to varying combinations of 60 μmol·m^-2·s^-1 of 405 nm LED and serial 10-fold dilutions of formalin (from 10 to 100 ppm) for 15, 30, 45, and 60 min. Surviving M. avidus were counted using a hemocytometer. For in vivo test, flounder acclimatized at 11-12 practical salinity unit (psu) were challenged with 2×10⁶ cells/ml of M. avidus by immersion method for 1 h. Then, fish were moved and divided into four groups; "F" group, treated with formalin at 50 ppm; "L" group, treated with 60 μmol·m^-2·s^-1 of 405 nm LED; "C" group, treated with combination of the two methods; and the control group. After treatment for 30 min, fish were transferred to new tanks (salinity = 11-12 psu) and observed for 3 weeks. As a result, illumination of 405 nm LED at 60 μmol·m^-2·s^-1 killed 100% of M. avidus after 12 h, while 67% and 90% of the scuticociliate died at 20 and 40 μmol·m^-2·s^-1, respectively, after 24 h exposure. One hundred percent of M. avidus was killed at 90, 80, 80 and 70 ppm after exposure to formalin for 15, 30, 45 and 60 min, respectively. However, combined method (e.g., 60 μmol·m^-2·s^-1 of 405 nm-LED plus 50 ppm formalin) killed the parasite within 30 min. From in vivo test, similarly, survival rates of fish challenged with M. avidus were 100%, 43%, 29% and 0% in the C, F, L, and control groups, respectively. Results obtained in this study demonstrates that the combined treatment method has clear synergistic effect on scuticociliatosis in fish.

• Tuberculosis and Respiratory Diseases
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• v.51 no.3
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• pp.211-223
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• 2001

Background : Rhinovirus infection of the airways results in increased permeability of the airway vascular endothelium with the influx of plasma proteins, including lipids such as LDL. In vitro studies on the effect of oxLDL on leukocytes has shown many pro inflammatory effects on multiple leukocytes. We hypothesized that oxLDL is one mechanism for recruiting granulocytes to the airways during a RV infection. Therefore, chemotaxis and transendothelial migration, in response to nLDL, was determined for these granulocytes. Methods : nLDL was oxidized with 5mM Cu2S04 for 20-24 hours. 3-5 105 cells were loaded into the Transwell filter while the chemotatic agonists were placed in the lower well for chemotaxis. Confluent monolayers on HPMEC were grown on Transwell filters for transendothelial migration. The filters were washed and eosinophils and neutrophils loaded on to the filter with the chemotatic agonist was were placed in the lower well. The wells were incubated for 3 hours. The number of migrating cells was counted on a hemocytometer. Results : OxLDL, but not nLDL, is chemotatic for eosinophils and neutrophils. The level of granulocytes chemotaxis was dependent on both the concentration of LDL and its degree of oxidation. OxLDL stimulates eosinophil and neutrophils migration across HPMEC monolayers (+/-IL-$1{\beta}$ preactivation) in a dose dependent manner. Conclusion : Increased vascular permeability during a RV infection may lead to the influx and oxidation of LDL. The resulting oxLDL. is one possible mechanism for the recruitment of neutrophils and eosinophils to the airway interstitial matrix. Once in the airways, granulocytes can further interact with oxLDL to promote airway inflammation.

• Journal of Periodontal and Implant Science
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• v.26 no.2
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• pp.469-490
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• 1996

The initial events required for periodontal regeneration is the attachment, spreading, and proliferation of appropriated cells at the healing sites. These have been reported that minocycline stimulates the attachment of periodontal ligament cells, and also $TGF-{\beta}1$ enhances the proliferation of periodontal ligament cells. The purpose of the present study was to evaluate the effects of $TGF-{\beta}1$ on the cellular activity of minocycline treated human periodontal ligament cells. Periodontal ligament cells were obtained from the explants of healthy periodontal ligaments of extracted 3rd molars or premolar teeth extracted from the patients for orthodontic treatment. The cells were cultured in minimal essential medium(${\alpha}-MEM$) supplemented with 10.000units/ml penicillin, $10,000{\mu}g/ml$ streptomycin and 10% FBS(fetal bovine serum) at $37^{\circ}C$ in a humidified atmosphere of 5% carbon dioxide and the 5th to the 8th passages of the cells were used. To evaluate the effect of minocycline on cell attachment, the cells were seeded at a cell density of $1.5{\times}10^4$ cells/well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After trypsinization, the cells were counted with hemocytometer and were taken photographs for observation of cellular morphology. To evaluate the effect of $TGF-{\beta}1$ on minocycline-pretreated periodontal ligament cells, the cells were seeded at a cell density of $1{\times}10^4$ cells/ well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After incubation, 1 and 10ng/ml of $rh-TGF-{\beta}1$ were also added to the each well and incubated for 1 and 2 days, respectively. Then, MTT assay, DNA synthesis($^3H-thymidine\;assay$), and protein and collagen assay(3H-proline assay) were carried out. In the MIT assay, after 200ul MTT solutionlconeentration of 5mg/ml) were added to the each well of the 24-well plates and incubated for 3 hours, and 200 ul DMSO were added so as to dissolve insoluble blue formazan crystals which was formed in incubated period. Then it read plates on a ELISA reader. For mitogenic assay, 1 uCi/ml $^3H-thymidine$ was added to each well for the final 2 hours of the incubation periods. After labeling, the wells were washed 3 times with ice cold PBS and 4 times with 5% TCA to remove unincorporated label and precipitate the cellular DNA. DNA, with the incorporated $^3H-thymidine$, was solubilized with 500 ul of 0.1% NaOH/0.1% SDS. A 250 ul aliquot was removed from each well and placed in a scintillation vial with 4ml of scintillation cocktail. Using an liguid scintillation counter, counts per minute(CPM) were determined for each samples. 3 uCi/ml $^3H-proline$ was added to each well for the final 4 hours of the incubation periods and total protein and percent collagen synthesis were carried out. The results indicate that minocycline treated group with $100{\mu}g/ml$ concentration for 1.5 hours significantly increased than that of control in cell attachment, and cell process is also evident compared with that of control in cell morphology, and the cellular activity and DNA synthesis rate of cells treated minocycline and $TGF-{\beta}1$ significantly increased than that of control values, but were below to values of the $TGF-{\beta}1$ only treated group in MIT assay and $^3H-thymidine\;assay$, and the total protein synthesis of minocycline and $TGF-{\beta}1$ treated group also significantly increased than that of control values, but the percent collagen synthesis of tested group significantly decreased to compared with control. On the above the findings, the tested group of minocycline and $TGF-{\beta}1$ did not increase the effect on the cell activity than $TGF-{\beta}1$ only tested group and the tested group of minocycline inhibited cell activity. This results indicate that minocycline was effective on cell attachment in early stage, but it is harmful to cell activity, that inhibitory effect of minocycline was compensated with stimulatory effect of $TGF-{\beta}1$.

• Tuberculosis and Respiratory Diseases
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• v.41 no.6
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• pp.604-615
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• 1994

Background: Analysis of cells in bronchoalveolar lavage(BAL) fluid had been used to predict the histologic changes of the bronchioles and alveoli in patients with interstitial lung diseases(ILD). Definitive diagnosis can be a1so made in some cases of ILD, such as histiocytosis. However, there are a few data of the cellular components in BAL fluid in normal Korean individuals and in patients with ILD. In order to evaluate the role of the cellular analysis of BAL fluid in prediction of alveolitis and differential diagnosis among ILDs, we compared the cellular components in BAL fluid from 50 normal individuals and 86 ILD patients. Method: BAL was performed by instillation and retrievement of normal saline with fiberoptic bronchoscopy. The cell number was counted by Hemocytometer. Differential count was done up to 500 cells on slides prepared by Diff-Quik stain and non-specific esterase stain. We compared the recovery rate(RR), cell numbers(CN), and percentages of each cellular components(CP). Results: The results were as follows: 1) There was no difference in RR, CN and CP between the normal smoker group and normal non-smoker group. 2) Total cell numbers recoverd in BAL fluid increased in collagen vascular diseases(CVD), hypersensitivity pneumonitis(HP), idiopathic pulmonary fibrosis(IPF), and miliary tuberculosis(Mil TBC) groups. 3) The percentage of lymphocytes increased in HP, IPF and Mil TBC groups. Macrophage percentages increased in HP, IPF, and Mil TBC groups. Neutrophil percentages were increased in CVD, HP, IPF and Mil TBC groups. Eosinophil percentages were increased in HP, IPF and Mil TBC groups. The numbers of each cells showed same findings as the percentages did. Conclusion: The analysis of cellular components of BAL fluid can predict the presence of alveolitis in many cases of ILDs. However, It was not helpful in differential diagnosis among ILDs.